Cellular Integration of Information in the Detection and Response to Epithelial Damage

上皮损伤检测和反应中的细胞信息整合

• 批准号: 9893174
• 负责人: M. Shane Hutson
• 金额: $ 21.5万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893174
• 关键词: Address; Adopted; Animals; Area; Behavior; Calcium; Calcium Signaling; Caliber; Cell Culture System; Cell Proliferation; Cell model; Cells; Cellular biology; Communication; Conflict (Psychology); Confusion; Data; Detection; Development; Diffuse; Distal; Drosophila genus; Epithelial; Epithelial Cells; Flare; Frequencies; Gap Junctions; Genetic; Genetic Transcription; Hour; Image; Individual; Knowledge; Lead; Literature; Location; MAPK8 gene; Malignant Neoplasms; Mechanics; Mediating; Membrane; Modeling; Molecular; Organism; Pathology; Pathway interactions; Pattern; Phylogeny; Process; Proliferating; Publishing; Pupa; Radial; Reporting; Role; Running; Side; Signal Transduction; Spatial Behavior; Stereotyping; System; Tissues; Work; Wound Healing; calcium indicator; cell behavior; cell injury; experience; extracellular; genetic manipulation; in vivo; mathematical model; migration; millisecond; programs; receptor; recruit; repaired; response; skills; tool; wound

Project Summary When an epithelial tissue is wounded, the first cellular response is a dramatic increase in cytosolic calcium levels, beginning immediately upon tissue damaged. This calcium increase is observed not just in cells at the wound margin but in large domain of cells surrounding the wound. With the advent of genetically encoded calcium indicators like GCaMP, this wound-induced calcium response has been observed in living organisms across the animal kingdom, yet conflicting mechanisms have been proposed to explain the induction of calcium, such that there is no sense of a conserved fundamental process. Our collaborative team of biophysicists and developmental geneticists recently published work identifying a major underlying obstacle: there are several contemporaneous mechanisms underlying the wound-induced calcium response, which we have been able to tease apart with our combination of highly quantitative approaches and our genetic manipulations. Without understanding the multiple mechanisms inducing calcium responses, it has been impossible to draw parallels across the literature and across wounding models, and it has impossible to fully block calcium responses to analyze the downstream consequences for wound healing. Our analysis tools identified stereotyped calcium responses, with different oscillatory patterns or signatures evident at different radial distances from the wound. We hypothesize that these patterned calcium responses inform the cell about its distance from the wound and determine its downstream cell behaviors. Like the calcium responses, wound-induced cell behaviors and transcriptional identities are patterned according to distance from the wound, with migratory cells and JNK signaling near the margin and proliferative cells and JAK-STAT signaling in a more distal ring. In the first Aim, we investigate the mechanisms of how calcium signaling is initiated in each of these patterns, working both with individual pathways and developing mathematical models for how these calcium patterns are integrated. In the second Aim, we perturb specific aspects of calcium patterns and ask how downstream cell behavior and identity are altered. We are able to achieve these Aims because of our unique collaborative skill-set, and because we have developed an unparalleled wounding model that allows genetic manipulation with high temporal control on one side of the wound only. Because it is internally controlled, comparing the two sides allows precise quantification and detection of even small changes, in both calcium signaling and wound-induced cell behaviors. At the completion of this project, we expect to have generated a high-precision model of how cells detect tissue damage at a distance, and how they interpret this information to select a spatially-appropriate repair program. This fundamental knowledge will be important to many areas of cell biology, to wound-healing studies, and to pathologies like cancer where wound-healing programs are inappropriately activated.

项目概要 当上皮组织受伤时，第一个细胞反应是细胞质急剧增加 钙水平，在组织受损后立即开始。这种钙的增加不仅出现在 细胞位于伤口边缘，但在伤口周围的大范围细胞中。随着基因技术的出现 编码钙指示剂，如 GCaMP，这种伤口诱导的钙反应已在活体中观察到 整个动物界的有机体，但有人提出了相互矛盾的机制来解释这一现象 钙的诱导，这样就没有保守的基本过程的意义。我们的协作团队 生物物理学家和发育遗传学家最近发表的工作确定了一个主要的潜在障碍： 伤口引起的钙反应有几种同时发生的机制，我们 已经能够通过我们高度定量的方法和我们的遗传方法的结合来梳理 操纵。在不了解诱导钙反应的多种机制的情况下， 不可能在文献和伤害模型之间进行比较，而且也不可能完全 阻断钙反应以分析伤口愈合的下游后果。 我们的分析工具识别出具有不同振荡模式或不同模式的钙反应 在距伤口不同径向距离处有明显的特征。我们假设这些图案化的钙 反应告知细胞其与伤口的距离，并确定其下游细胞的行为。喜欢 钙反应、伤口诱导的细胞行为和转录特性的模式根据 距离伤口较远，边缘附近有迁移细胞和 JNK 信号传导，并且有增殖细胞 JAK-STAT 信号传导在更远端的环中。在第一个目标中，我们研究了钙如何 信号传导在每种模式中启动，与单独的途径一起工作并发展 这些钙模式如何整合的数学模型。在第二个目标中，我们扰乱特定的 钙模式的各个方面，并询问下游细胞的行为和身份是如何改变的。我们能够 实现这些目标是因为我们独特的协作技能，并且因为我们开发了 无与伦比的伤害模型，允许在一侧进行高时间控制的基因操作 仅伤口。由于它是内部控制的，因此比较两侧可以进行精确的量化和 检测钙信号传导和伤口诱导的细胞行为的微小变化。 在该项目完成后，我们期望生成一个高精度模型，说明细胞如何 检测远处的组织损伤，以及他们如何解释这些信息以选择空间合适的 修复程序。这一基础知识对于细胞生物学的许多领域、伤口愈合都很重要 研究，以及癌症等伤口愈合程序被不当激活的病理学。